CD8+HLA-DR+ T cells are increased in patients with severe aplastic anemia

Xing,Limin; Liu,Chunyan; Fu,Rong; Wang,Huaquan; Wang,Jun; Liu,Xiao; Feng,Le; Li,Lijuan; Liu,Hui; Wang,Honglei; Zhang,Tian; Shao,Zonghong

doi:10.3892/mmr.2014.2344

CD8+HLA-DR+ T cells are increased in patients with severe aplastic anemia

Authors:
- Limin Xing
- Chunyan Liu
- Rong Fu
- Huaquan Wang
- Jun Wang
- Xiao Liu
- Le Feng
- Lijuan Li
- Hui Liu
- Honglei Wang
- Tian Zhang
- Zonghong Shao
View Affiliations

Affiliations: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, P.R. China
Published online on: June 18, 2014 https://doi.org/10.3892/mmr.2014.2344
Pages: 1252-1258

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate the number and function of CD8+HLA-DR+ cells, which are considered to be activated cytotoxic T lymphocytes (CTLs), in peripheral blood to further examine the pathogenesis of severe aplastic anemia (SAA). Thirty-eight patients with SAA were included in the present study. Patients were screened for paroxysmal nocturnal hemoglobinuria by flow cytometry using anti-CD55 and anti-CD59 antibodies. The number of CD8+HLA-DR+ T cells was measured by three-color flow cytometry using anti-CD8-peridinin chlorophyll, anti-CD3-fluorescein isothiocyanate (FITC) and anti-HLA-DR-FITC antibodies. The expression of perforin, granzyme B, tumor necrosis factor-β (TNF-β) and FasL in CD8+HLA-DR+ T cells was detected by flow cytometry with the appropriate monoclonal antibodies. Total RNA was prepared from purified CD8+HLA-DR+ cells of healthy controls and SAA patients, and then polymerase chain reaction (PCR) was performed. Apoptosis of CD8+HLA-DR+ cells was detected by flow cytometry following staining with Annexin V. The proportion of CD8+HLA-DR+ T cells was analyzed by flow cytometry in peripheral blood and was identified to be significantly higher in untreated SAA than in remission patients and in the controls. The expression of perforin, granzyme B, TNF-β and FasL in CD8+HLA-DR+ T cells was analyzed by flow cytometry and PCR, which revealed increased expression in the untreated SAA group compared with that in the control group. Furthermore, the apoptosis of CD3- bone marrow cells from normal individuals was enhanced following co-culture with CD8+HLA-DR+ T cells from untreated SAA patients. In conclusion, the present study demonstrated that CD8+HLA-DR+ T cells may contribute to bone marrow failure in SAA.

View Figures

View References

1	Young NS, Bacigalupo A and Marsh JC: Aplastic anemia: pathophysiology and treatment. Biol Blood Marrow Transplant. 16(Suppl 1): S119–S125. 2010. View Article : Google Scholar : PubMed/NCBI
2	Bacigalupo A: Aplastic anemia: pathogenesis and treatment. Hematology Am Soc Hematol Educ Program. 2007:23–28. 2007. View Article : Google Scholar
3	Marsh JC, Ball SE, Cavenagh J, et al: Guidelines for the diagnosis and management of aplastic anaemia. Br J Haematol. 147:43–70. 2009. View Article : Google Scholar
4	Keckler MS: Dodging the CTL response: viral evasion of Fas and granzyme induced apoptosis. Front Biosci. 12:725–732. 2007. View Article : Google Scholar : PubMed/NCBI
5	Klajman A, Drucker I and Mekori J: The importance of HLA-DR in autoimmune diseases. Isr J Med Sci. 26:691–692. 1990.PubMed/NCBI
6	Jeong DC, Chung NG, Cho B, et al: Long-term outcome after immunosuppressive therapy with horse or rabbit antithymocyte globulin and cyclosporine for severe aplastic anemia in children. Haematologica. 99:664–671. 2014. View Article : Google Scholar
7	Shao ZH: The research of aplastic anemia. Basic Clin Med. 27:233–237. 2007.(In Chinese).
8	Zonghong S, Meifeng T, Huaquan W, et al: Circulating myeloid dendritic cells are increased in individuals with severe aplastic anemia. Int J Hematol. 93:156–162. 2011. View Article : Google Scholar : PubMed/NCBI
9	Solomou EE, Keyvanfar K and Young NS: T-bet, a Th1 transcription factor, is up-regulated in T cells from patients with aplastic anemia. Blood. 107:3983–3991. 2006. View Article : Google Scholar : PubMed/NCBI
10	He GS, Shao ZH and He H: Quantitative and functional changes of T helper cell subsets in the bone marrow of severe aplastic anemia patients. Zhonghua Xue Ye Xue Za Zhi. 25:613–616. 2004.(In Chinese).
11	Li ZS, Shao ZH, Fu R, et al: Percentages and functions of natural killer cell subsets in peripheral blood of patients with severe aplastic anemia. Zhonghua Yi Xue Za Zhi. 91:1084–1087. 2011.(In Chinese).
12	Passweg JR and Marsh JC: Aplastic anemia: first-line treatment by immunosuppression and sibling marrow transplantation. Hematology Am Soc Hematol Educ Program. 2010:36–42. 2010. View Article : Google Scholar : PubMed/NCBI
13	Locasciulli A, Oneto R, Bacigalupo A, et al: Outcome of patients with acquired aplastic anemia given first line bone marrow transplantation or immunosuppressive treatment in the last decade: a report from the European Group for Blood and Marrow Transplantation (EBMT). Haematologica. 92:11–18. 2007. View Article : Google Scholar
14	Hattori M, Terasawa T, Tsushita K, et al: The status of antithymocyte globulin therapy for adult patients in Japan: retrospective analysis of a nationwide survey. Int J Hematol. 87:48–55. 2008. View Article : Google Scholar
15	Young NS: Pathophysiologic mechanisms in acquired aplastic anemia. Hematology Am Soc Hematol Educ Program. 2006:72–77. 2006. View Article : Google Scholar
16	Mekmullica J, Brouwers P, Charurat M, et al: Early immunological predictors of neurodevelopmental outcomes in HIV-infected children. Clin Infect Dis. 48:338–346. 2009. View Article : Google Scholar : PubMed/NCBI
17	Viallard JF, Blanco P, André M, et al: CD8⁺HLA-DR⁺ T lymphocytes are increased in common variable immunodeficiency patients with impaired memory B-cell differentiation. Clin Immunol. 119:51–58. 2006.
18	Pipkin ME and Lieberman J: Delivering the kiss of death: progress on understanding how perforin works. Curr Opin Immunol. 19:301–308. 2007. View Article : Google Scholar : PubMed/NCBI
19	Pardo J, Aguilo JI, Anel A, et al: The biology of cytotoxic cell granule exocytosis pathway: granzymes have evolved to induce cell death and inflammation. Microbes Infect. 11:452–459. 2009. View Article : Google Scholar : PubMed/NCBI
20	Blanco P, Pitard V, Viallard JF, et al: Increase in activated CD8⁺ T lymphocytes expressing perforin and granzyme B correlates with disease activity in patients with systemic lupus erythematosus. Arthritis Rheum. 52:201–211. 2005.
21	Prpić Massari L, Kastelan M, Gruber F, et al: Perforin expression in peripheral blood lymphocytes and skin-infiltrating cells in patients with lichen planus. Br J Dermatol. 151:433–439. 2004.PubMed/NCBI
22	Lettau M, Paulsen M, Kabelitz D and Janssen O: FasL expression and reverse signalling. Results Probl Cell Differ. 49:49–61. 2009. View Article : Google Scholar
23	Nadeau KC, Callejas A, Wong WB, et al: Idiopathic neutropenia of childhood is associated with Fas/FasL expression. Clin Immunol. 129:438–447. 2008. View Article : Google Scholar : PubMed/NCBI
24	Takaoka Y, Abe Y, Haraguchi R and Kito K: Lymphotoxin (TNF-beta). Nihon Rinsho. 68(Suppl 7): 93–95. 2010.(In Japanese).
25	Kamio T, Ito E, Ohara A, et al: Relapse of aplastic anemia in children after immunosuppressive therapy: a report from the Japan Childhood Aplastic Anemia Study Group. Haematologica. 96:814–819. 2011. View Article : Google Scholar
26	Bacigalupo A, Bruno B, Saracco P, et al: Antilymphocyte globulin, cyclosporine, prednisolone, and granulocyte colony-stimulating factor for severe aplastic anemia: an update of the GITMO/EBMT study on 100 patients. European Group for Blood and Marrow Transplantation (EBMT) Working Party on Severe Aplastic Anemia and the Gruppo Italiano Trapianti di Midolio Osseo (GITMO). Blood. 95:1931–1934. 2000.